Distribution and properties of lactate dehydrogenase isoenzymes in red and white muscle of freshwater fish

Abstract

The distribution and kinetics of LDH isoenzymes in red and white muscles of 5 species of salmonids, 4 species of cyprinids and one coregonid species were studied. In all species the white muscles are characterized by the occurrence of only the most cathodic isoenzymes, or groups of isoenzymes. The red muscles contained either the full set of isoenzymes (cyprinids) or a selection in which the anodic forms dominated (salmonids, coregonid). The most striking difference between the two types of muscle was met inCoregonus sp. The temperature profiles of pyruvate affinity are similar in all species of fish studied. On the other hand, Km(pyr) values and degree of pyruvate inhibition are closely related and vary greatly with temperature, with the taxonomic position (and thus biology) of the species, and with electrophoresic mobility of the isoenzyme. Highest affinity and strongest inhibition occurred in the anodic (H4) isoenzymes of cyprinids at low temperature; lowest affinity and zero inhibition in the cathodic isoenzymes (Mα4 → Mβ4) of salmonids and coregonids at high temperature. In salmonids the more recently duplicated loci of the M-group of isoenzymes possess identical Km values at all temperatures, whereas the two older M and H loci differ greatly in this respect. Thus the more recent duplication of LDH loci in salmonids and coregonids may be seen as a mechanism by which the tetramers required for LDH activity can be constructed from more closely related subunits than are provided by the older M and H loci.Some problems in connection with the determination of the kinetic constants of the lactate oxidase reaction are discussed and it is suggested that an alkaline, pyruvate trapping system provides conditions which are more realistic than those of other assay systems. The Km(lactate) values found are in the biological range and, at 20°C, provide further circumstantial evidence that the red muscles of fish should be capable of oxidizing the lactate produced by the white muscles during strenuous exercise. At 4°C the Km(lactate) values are abnormally high in all muscle preparations and thus are not correlated with the Km(pyruvate) values which are lowest at this temperature.